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Analysis of gene expression profile and T cell and B cell receptor diversities in the fetuses and trafficked maternal cells following murine in-utero transplantation of semi-allogenic bone marrow-derived hematopoietic stem cells

ABSTRACT: We investigated the immune responses in the fetuses and trafficked maternal cells following in-utero transplantation (IUT) of maternal (mIUT) and paternal (pIUT) bone marrow donor mononuclear cells (MNC). Fetal cells and trafficked maternal cells were isolated from the 1 week old neonates by magnetic and FACS sorting and subjected to bulk RNA-sequencing. The gene expression profile in maternal and recipient cells of mIUT and pIUT show enriched clusters for RNA and protein metabolism, hemopoiesis and immune system development. In recipients of DT+ pIUT recipient clusters additionally represented T-cell regulation. Upregulated pIUT and DT+pIUT maternal clusters represented mitogen-activated protein kinases (MAPK) cascades, T-cell activation, and immune system development. All groups shared common genes represented for cytokine stimulus response, immune system regulation, B-cell mediated immunity and adaptive immune response. Analysis of receptor clonotypes reveals, the retrieval frequency of maternal-derived clonotypes increased and clonotypes diversity was reduced following mIUT and pIUT compared to uninjected controls. In DT+pIUT, decreased maternal clonotype retrieval and increased diversity towards baseline. In contrast, mIUT and pIUT recipient-derived clonotypes showed increased diversity while decreased frequency in DT+pIUT recipients. Maternal-derived top 5 clonotypes in each group were expanded with IUT and diminished with DT+, while recipient-derived top 5 clonotypes were most abundant in uninjected pups. We observed a large number of public maternal-derived clonotypes between DT+pIUT and controls, and a substantial number of public recipient-derived clonotypes between mIUT, pIUT and DT+pIUT. Complementarity-determining region 3 (CDR3) of naïve and antigen-experienced clonotypes are longer and shorter respectively, reflecting antigen driven selection. We also observed uniqueness in V/J-segment usage between uninjected control, mIUT and pIUT clonotypes. Our data indicate that both maternal and recipient immune cells respond to donor cells and that only maternal TCR and BCR diversity is affected by maternal DC depletion.

ORGANISM(S): Mus musculus

PROVIDER: GSE195771 | GEO | 2022/02/03

REPOSITORIES: GEO

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Project description:Tumor-Infiltrating Lymphocytes (TILs) are composed by several immune subpopulations including NK, NKT, Tab, Tgd and B cells. Although some of these cells can have antitumoral activities, there is also a large number of cells which are not tumor-specific,known as bystander cells. These cells may not have a direct role in the antitumoral response but could lead to a modulation of the immune response after time. Recent studies have described fatal encephalitis and myocarditis in cancer patients after immune checkpoint therapies, in which EBV-specific T cells may be involved. Since EBV is able to infect B cells and epithelial cells, both present in the primary tumor site, the first events of these fatal responses could be a consequence of a dysregulation of the immune system in the tumor microenvironment, although this have not been deeply studied. Thus, the main objectives of this study are (i) to use EBV-transformed B cells from the tumor site as a model to study the tumor-infiltrating EBV+ B cells and (ii) to use these EBV-derived B cells to study the relationship with autologous T cells isolated from the tumor site. T cells from TILs have been isolated from a triple negative breast cancer patient before neoadjuvancy and were expanded directly from the biopsy. A Patient-Derived Xenograft (PDX) generated from the breast tumor, progressed into a lymphocytic neoplasm. EBVtransformed B cells were obtained from this PDX (PDX-derived B cells). T Cell Receptor high-throughput sequencing has been used to monitor the clonotypes present in the different samples. We have found 43 clonotypes infiltrating the PDX-562 tissue and 18 of them were also present in the human biopsy or in the biopsy-derived T cell cultures. 5 of these sequences were found in one of the biopsy-derived T cells analyzed. Thus, PDXderived B cells have been used as antigen presenting cells to expand these cells. Few clonotypes were obtained after expansions, although the analysis of the middle CDR3 have revealed a similar pattern with the 18 sequences from the PDX sample. Furthermore, peptides presented by HLA have been eluted from PDX-derived B cells, followed by MS analysis. Peptides obtained derived from 31 different proteins and >40% are cancerrelated in terms of expression.

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Analysis of gene expression profile and T cell and B cell receptor diversities in the fetuses and trafficked maternal cells following murine in-utero transplantation of semi-allogenic bone marrow-derived hematopoietic stem cells

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